Optogenetics for light control of biological systems

Nature Reviews Methods Primers, Год журнала: 2022, Номер 2(1)

Опубликована: Июль 21, 2022

Язык: Английский

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Base editing rescue of spinal muscular atrophy in cells and in mice

Science, Год журнала: 2023, Номер 380(6642)

Опубликована: Март 30, 2023

Spinal muscular atrophy (SMA), the leading genetic cause of infant mortality, arises from survival motor neuron (SMN) protein insufficiency resulting

Язык: Английский

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Two-photon calcium imaging of neuronal activity

Nature Reviews Methods Primers, Год журнала: 2022, Номер 2(1)

Опубликована: Сен. 1, 2022

Язык: Английский

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Optimal trade-off control in machine learning–based library design, with application to adeno-associated virus (AAV) for gene therapy

Science Advances, Год журнала: 2024, Номер 10(4)

Опубликована: Янв. 24, 2024

Adeno-associated viruses (AAVs) hold tremendous promise as delivery vectors for gene therapies. AAVs have been successfully engineered—for instance, more efficient and/or cell-specific to numerous tissues—by creating large, diverse starting libraries and selecting desired properties. However, these often contain a high proportion of variants unable assemble or package their genomes, prerequisite any goal. Here, we present showcase machine learning (ML) method designing AAV peptide insertion that achieve fivefold higher packaging fitness than the standard NNK library with negligible reduction in diversity. To demonstrate our ML-designed library’s utility downstream engineering goals, show it yields approximately 10-fold successful after selection infection human brain tissue, leading promising glial-specific variant. Moreover, design approach can be applied other types beyond.

Язык: Английский

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A high-fidelity CRISPR-Cas13 system improves abnormalities associated with C9ORF72-linked ALS/FTD

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Янв. 8, 2025

Abstract An abnormal expansion of a GGGGCC (G 4 C 2 ) hexanucleotide repeat in the C9ORF72 gene is most common genetic cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two debilitating neurodegenerative disorders driven part by gain-of-function mechanisms involving transcribed forms expansion. By utilizing Cas13 variant with reduced collateral effects, we develop here high-fidelity RNA-targeting CRISPR-based system for C9ORF72-linked ALS/FTD. When delivered to brain transgenic rodent model, this Cas13-based platform curbed expression G repeat-containing RNA without affecting normal levels, which turn decreased formation foci, production dipeptide protein, reversed transcriptional deficits. This possessed improved transcriptome-wide specificity compared its native form mediated targeting motor neuron-like cells derived from patient ALS. These results lay foundation implementation CRISPR technologies

Язык: Английский

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Dual-targeting CRISPR-CasRx reduces C9orf72 ALS/FTD sense and antisense repeat RNAs in vitro and in vivo

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Янв. 8, 2025

Abstract The most common genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) is an intronic G 4 C 2 repeat expansion in C9orf72 . repeats undergo bidirectional transcription to produce sense antisense RNA species, which are translated into dipeptide proteins (DPRs). As toxicity has been associated with both repeat-derived DPRs, targeting strands may provide the effective therapeutic strategy. CRISPR-Cas13 systems mature their own guide arrays, allowing multiple species from a single construct. We show CRISPR-Cas13d variant CasRx effectively reduces overexpressed transcripts DPRs HEK cells. In patient-derived iPSC-neuron lines, CRISPR-CasRx endogenous RNAs protects against glutamate-induced excitotoxicity. AAV delivery two distinct mouse models significantly reduced repeat-containing transcripts. This highlights potential RNA-targeting CRISPR as therapeutics for ALS/FTD.

Язык: Английский

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Systemic administration of novel engineered AAV capsids facilitates enhanced transgene expression in the macaque CNS

Med, Год журнала: 2022, Номер 4(1), С. 31 - 50.e8

Опубликована: Ноя. 22, 2022

Adeno-associated virus (AAV) vectors are a promising vehicle for noninvasive gene delivery to the central nervous system via intravenous infusion. However, naturally occurring serotypes have limited ability transduce brain, and translating engineered capsids from mice nonhuman primates has proved challenging.In this study, we use an mRNA-based directed-evolution strategy in multiple strains of as well de novo selection cynomolgus macaques identify families with increased potency brain decreased tropism liver.We compare transgene expression capabilities several show that while some our novel macaque-derived variants significantly outperform AAV9 transducing macaque following systemic administration, mouse-derived variants-both those identified study reported by other groups-universally do not.Together, results work introduce class primate-derived AAV therapeutic potential highlight critical need using appropriate animal models both evaluate AAVs intended human system.This was funded primarily through anonymous philanthropic gift P.C.S. lab at Broad Institute MIT Harvard grant Howard Hughes Medical

Язык: Английский

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Advances in AAV technology for delivering genetically encoded cargo to the nonhuman primate nervous system

Current Research in Neurobiology, Год журнала: 2023, Номер 4, С. 100086 - 100086

Опубликована: Янв. 1, 2023

Modern neuroscience approaches including optogenetics, calcium imaging, and other genetic manipulations have facilitated our ability to dissect specific circuits in rodent models study their role neurological disease. These regularly use viral vectors deliver cargo (e.g., opsins) tissues genetically-engineered rodents achieve cell-type specificity. However, the translatability of these models, cross-species validation identified targets, translational efficacy potential therapeutics larger animal like nonhuman primates remains difficult due lack efficient primate vectors. A refined understanding nervous system promises insights that can guide development treatments for neurodegenerative diseases. Here, we outline recent advances adeno-associated optimized primates. tools promise help open new avenues further brain.

Язык: Английский

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The menace of severe adverse events and deaths associated with viral gene therapy and its potential solution

Medicinal Research Reviews, Год журнала: 2024, Номер 44(5), С. 2112 - 2193

Опубликована: Март 28, 2024

Abstract Over the past decade, in vivo gene replacement therapy has significantly advanced, resulting market approval of numerous therapeutics predominantly relying on adeno‐associated viral vectors (AAV). While have undeniably addressed several critical healthcare challenges, their clinical application unveiled a range limitations and safety concerns. This review highlights emerging challenges field therapy. At first, we discuss both role biological barriers with focus AAVs, current landscape human We delineate advantages disadvantages AAVs as delivery vehicles, mostly from perspective (hepatotoxicity, cardiotoxicity, neurotoxicity, inflammatory responses etc.), outline mechanisms adverse events response to AAV. Contribution every aspect AAV (genomic structure, capsid proteins) host injected is considered substantiated by basic, translational studies. The updated evaluation recent trials medical experience clearly shows risks that sometimes overshadow hopes for curing hereditary disease. last, set established new molecular nanotechnology tools approaches are provided potential solutions mitigating or eliminating side effects. increasing number severe reactions and, sadly deaths, demands decisive actions resolve issue immune extremely high doses used In these various strategies under development, including aimed at augmenting characteristics others focused creating secure efficacious non‐viral vectors. comprehensive offers an overarching present state utilizing

Язык: Английский

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Gene therapy clinical trials worldwide to 2023—an update

The Journal of Gene Medicine, Год журнала: 2024, Номер 26(8)

Опубликована: Авг. 1, 2024

Abstract To date, 3,900 gene therapy clinical trials have been completed, are ongoing or approved worldwide. Our database brings together global information on activity from trial databases, official agency sources, published literature, conference presentations and posters kindly provided to us by individual investigators sponsors. This review presents our analysis of that, the best knowledge, being performed As March 2023 update, we entries undertaken in 46 countries. We analyzed geographical distribution trials, disease indications (or other reasons) for proportions which different vector types used, genes transferred. Details analyses presented, searchable The Journal Gene Medicine Therapy Clinical Trials Worldwide website at https://a873679.fmphost.com/fmi/webd/GTCT . also provide an overview progress made around world, discuss key trends since previous review, namely unprecedented increase activity, including implementation genome editing technology with potential transform field moving forward.

Язык: Английский

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